Comparison of Four Human Papillomavirus Genotyping Methods: Next-generation Sequencing, INNO-LiPA, Electrochemical DNA Chip, and Nested-PCR
2018; 38(2): 139-146
Korean J Lab Med 2004; 24(4): 255-260
Published online August 1, 2004
Copyright © Korean Society for Laboratory Medicine.
Background :Apolipoprotein E (apoE) polymorphism is associated with the risk and the time of onset of Alzheimer’s disease and the risk of developing cardiovascular disease. Therefore, much interest in apoE genotyping has been focused both for epidemiological research and for the purpose of diagnosing dyslipoproteinemia or dementia. The aim of our study was to compare and evaluate three different methods for apoE genotyping. Method :We chose 10 samples each of six different apoE genotypes determined by INNO-LiPA ApoE Kit (INNOGENETICS, Gent, Belgium) used as the first comparison method. The other two were a fluorescence resonance energy transfer assay that used real-time PCR on the LightCycler instrument (Roche Diagnostics, Mannheim, Germany) and BioCore ApoE Kit (BioCore, Seoul, Korea) using the multiplex Amplification Refractory Mutation System (ARMS). Result :All six genotypes for apoE were clearly discernible with INNO-LiPA ApoE Kit, allelic discrimination with LightCycler, and BioCore ApoE Kit. We obtained a 100% concordance rate among the three methods. It took approximately 6.5 hours, 70 minutes, and 3 hours, respectively, but handson time took 3 hours, 45 minutes, and 1 hour. Conclusion :The INNO-LiPA allelic specific oligonucleotide probe method is accurate and reliable, but labor intensive and relatively time consuming. The LightCycler allelic discrimination method seems to be rapid, simple and accurate, suggesting that it may be used successfully for diagnostic purposes. The BioCore multiplex ARMS analysis is reliable, simple to perform, and less time consuming; this method also may be appropriate for determining the apoE genotypes in clinical laboratories.
Keywords: Apolipoprotein E, Alzheimer’s disease, Coronary artery disease, INNO-LiPA, Light-Cycler, Amplification Refractory Mutation System